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Ruxolitinib (INCB18424) Phosphate JAK1/2-remmer

Name: Ruxolitinib (INCB18424) Phosphate
Brand: Selleck Chemicals
SKU: S5243
Rating: 5 (182 reviews)

Cat.Nr.S5243

Ruxolitinib Fosfaat (INCB018424, INC424) is de fosfaatzoutvorm van Ruxolitinib. Ruxolitinib is de eerste krachtige, selectieve JAK1/2-remmer die de kliniek ingaat met een IC50 van 3,3 nM/2,8 nM in celvrije assays, >130-voudige selectiviteit voor JAK1/2 versus JAK3. Ruxolitinib doodt tumorcellen door toxische mitophagy. Ruxolitinib induceert autophagy en versterkt apoptosis.

Ruxolitinib (INCB18424) Phosphate JAK remmer Chemical Structure

Chemische structuur

Moleculair gewicht: 404.36

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Kwaliteitscontrole

Batch: Zuiverheid: 99.99%

99.99

Gerelateerde doelwitten	EGFR STAT Pim
Overige JAK Inhibitoren	BMS-986165 (Deucravacitinib) AZD1480 WP1066 Momelotinib (CYT387) Filgotinib (GLPG0634) AT9283 Gandotinib (LY2784544) Pacritinib TG101209 Cerdulatinib (PRT062070) hydrochloride

Celkweek, behandeling & werkconcentratie

Cellijnen	Assaytype	Concentratie	Incubatietijd	Activiteitsbeschrijving	PMID
Sf21	Function assay		1 hr	Inhibition of human JAK2 kinase domain expressed in Sf21 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, IC50=0.0028μM	22591402
Sf21	Function assay		1 hr	Inhibition of human JAK1 kinase domain expressed in Sf21 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, IC50=0.0033μM	22591402
Sf21	Function assay		1 hr	Inhibition of human TYK2 kinase domain expressed in Sf21 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, IC50=0.019μM	22591402
Sf21	Function assay		1 hr	Inhibition of human JAK3 kinase domain expressed in Sf21 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, IC50=0.428μM	22591402
TF1	Function assay		20 mins	Inhibition of JAK2 in human TF1 cells assessed as inhibition of EPO-induced STAT5 phosphorylation incubated for 20 mins prior to EPO-induction measured after 30 to 45 mins, EC50=0.012μM	22698084
TF1	Function assay		20 mins	Inhibition of JAK1 in human TF1 cells assessed as inhibition of IL6-induced STAT3 phosphorylation incubated for 20 mins prior to IL6-induction measured after 30 to 45 mins, EC50=0.024μM	22698084
SET2	Function assay			Inhibition of JAK2 V617F mutant in human SET2 cells assessed as reduction in STAT5 phosphorylation, IC50=0.00184μM	23061660
TF1	Function assay		30 mins	Inhibition of JAK2 in human TF1 cells assessed as reduction in STAT5 phosphorylation incubated for 30 mins in presence of human recombinant EPO, IC50=0.00685μM	23061660
T-cells	Function assay			Inhibition of JAK3/1 in human T cells expressing CD3 assessed as inhibition of IL2-stimulated STAT5a phosphorylation, IC50=0.023μM	23540648
T-cells	Function assay			Inhibition of JAK2/1 in human T cells expressing CD3 assessed as inhibition of IFNgamma-stimulated STAT1 phosphorylation, IC50=0.031μM	23540648
Sf9	Function assay		1 hr	Inhibition of human JAK2 (828-1132) expressed in baculovirus-infected Sf9 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, Ki=0.0001μM	23668484
Sf9	Function assay		1 hr	Inhibition of human JAK1 (837-1142) expressed in baculovirus-infected Sf9 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, Ki=0.0002μM	23668484
Sf9	Function assay		1 hr	Inhibition of human TYK2 (873-1187) expressed in baculovirus-infected Sf9 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, Ki=0.0005μM	23668484
Sf9	Function assay		1 hr	Inhibition of human JAK3 (781-1124) expressed in baculovirus-infected Sf9 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, Ki=0.0032μM	23668484
CD34+	Function assay		45 mins	Inhibition of JAK2 homodimer in human CD34+ cells spiked into human whole blood assessed as inhibition of EPO-induced STAT-5 phosphorylation preincubated for 45 mins followed by EPO addition measured after 15 mins by FACS analysis, IC50=0.677μM	24417533
BA/F3	Antiproliferative assay		72 hrs	Antiproliferative activity against mouse BA/F3 cells expressing TEL-JAK1 after 72 hrs by cell titer glo assay	26258521
BA/F3	Antiproliferative assay		72 hrs	Antiproliferative activity against mouse BA/F3 cells expressing TEL-JAK2 after 72 hrs by cell titer glo assay	26258521
TALL-1	Function assay	1 uM	3 hrs	Inhibition of JAK3 in human TALL-1 cells assessed as inhibition of IL-2 induced STAT5 phosphorylation at 1 uM preincubated for 3 hrs followed by IL-2 induction measured after 30 mins by immunoblotting	26258521
OCL-AML5	Function assay	1 uM	3 hrs	Inhibition of JAK2 in human OCL-AML5 cells assessed as inhibition of GM-CSF induced STAT5 phosphorylation at 1 uM preincubated for 3 hrs followed by GM-CSF induction measured after 30 mins by immunoblotting	26258521
CD34+	Function assay			Inhibition of JAK2 in human CD34+ cells assessed as inhibition of EPO-mediated cell proliferation, IC50=0.008μM	26927423
PBMC	Function assay			Inhibition of JAK1 in human PBMC cells assessed as inhibition of IL-6-induced MCP1 secretion, IC50=0.04μM	26927423
PBMC	Function assay			Inhibition IL-7-indcued STAT5 phosphorylation in human PBMC cells by flow cytometry, IC50=0.448μM	26927423
Sf21	Function assay		60 mins	Inhibition of human recombinant JAK2 expressed in Sf21 cells assessed as reduction in Ulight-CAGAGAIETDKEYYTVKD phosphorylation pre-incubated before substrate addition and measured after 60 mins by LANCE detection method, IC50=0.003μM	27137359
Sf21	Function assay		1 hr	Inhibition of recombinant human C-terminal 6His-tagged JAK2 (808 to end amino acids) expressed in Sf21 cells measured after 1 hr in presence of ATP by TR-FRET assay, IC50=0.0041μM	27555284
BaF3	Function assay			Inhibition of JAK2 V617F mutant expressed in mouse BaF3 cells cells assessed as reduction in cell viability, EC50=0.186μM	27555284
HEL 92.1.7	Antiproliferative assay		3 days	Antiproliferative activity against HEL 92.1.7 cells assessed as viable cells measured after 3 days by WST-1 assay, IC50=14.7μM	27555284
HCC827	Function assay	30 uM	24 hrs	Inhibition of JAK2 in human gefitinib-resistant HCC827 cells assessed as inhibition of STAT3 phosphorylation at Y705 site at 30 uM measured after 24 hrs by western blotting analysis	27555284
HCC827	Function assay	30 uM	24 hrs	Inhibition of JAK2 in wild-type human HCC827 cells assessed as inhibition of STAT3 phosphorylation at Y705 site at 30 uM measured after 24 hrs by western blotting analysis	27555284
HEL	Antiproliferative assay		48 hrs	Antiproliferative activity against HEL cells harboring JAK2 V617F mutant after 48 hrs by MTT assay, IC50=2.62μM	27774135
K562	Antiproliferative assay		48 hrs	Antiproliferative activity against human K562 cells after 48 hrs by MTT assay, IC50=10.3μM	27774135
MOLT4	Antiproliferative assay		48 hrs	Antiproliferative activity against human MOLT4 cells after 48 hrs by MTT assay, IC50=15.8μM	27774135
NCI-H23	Antiproliferative assay			Antiproliferative activity against human NCI-H23 cells harboring KRAS G12C mutant at	28038940
NCI-H358	Antiproliferative assay			Antiproliferative activity against human NCI-H358 cells harboring KRAS G12C mutant at	28038940
A673	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells	29435139
SK-N-MC	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells	29435139
NB1643	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells	29435139
LAN-5	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells	29435139
HeLa	Function assay	0.1 to 1 uM	1 hr	Inhibition of JAK2 in human HeLa cells assessed as reduction in STAT5 phosphorylation at 0.1 to 1 uM after 1 hr by immunoblot analysis	30243158
HeLa	Function assay	0.1 to 1 uM	1 hr	Inhibition of JAK2 in human HeLa cells assessed as increase in JAK2 phosphorylation at 0.1 to 1 uM after 1 hr by immunoblot analysis	30243158
Sf9	Function assay		1 hr	Inhibition of human JAK2 (828 to 1132 residues) expressed in baculovirus infected Sf9 insect cells using EQEDEPEGDYFEWLE as substrate after 1 hr by fluorescence assay, IC50=0.0028μM	30833158
Sf9	Function assay		1 hr	Inhibition of human JAK1 (837 to 1142 residues) expressed in baculovirus infected Sf9 insect cells using EQEDEPEGDYFEWLE as substrate after 1 hr by fluorescence assay, IC50=0.0033μM	30833158
HEL	Antiproliferative assay		48 hrs	Synergistic antiproliferative activity against HEL cells harboring JAK2 V617F mutant assessed as reduction in cell viability after 48 hrs in presence of SAHA by MTT assay, IC50=0.3μM	30901208
K562	Antiproliferative assay		48 hrs	Synergistic antiproliferative activity against human K562 cells assessed as reduction in cell viability after 48 hrs in presence of SAHA by MTT assay, IC50=1.03μM	30901208
MOLT4	Antiproliferative assay		48 hrs	Antiproliferative activity against human MOLT4 cells assessed as reduction in cell viability after 48 hrs by MTT assay, IC50=15.8μM	30901208
HEL	Antiproliferative assay		48 hrs	Antiproliferative activity against HEL cells harboring JAK2 V617F mutant assessed as reduction in cell viability after 48 hrs by MTT assay, IC50=18.6μM	30901208
K562	Antiproliferative assay		48 hrs	Antiproliferative activity against human K562 cells assessed as reduction in cell viability after 48 hrs by MTT assay, IC50=23.2μM	30901208
A549	Function assay	80 to 400 nM	6 hrs	Inhibition of HDAC2 in human A549 cells assessed as increase in acetyl histone H4 level at 80 to 400 nM after 6 hrs by Western blot analysis	30901208
A549	Function assay	80 to 400 nM	6 hrs	Inhibition of HDAC1 in human A549 cells assessed as increase in acetyl histone H4 level at 80 to 400 nM after 6 hrs by Western blot analysis	30901208
A549	Function assay	80 to 400 nM	6 hrs	Inhibition of HDAC3 in human A549 cells assessed as increase in acetyl histone H4 level at 80 to 400 nM after 6 hrs by Western blot analysis	30901208
A549	Function assay	80 to 400 nM	6 hrs	Inhibition of HDAC6 in human A549 cells assessed as increase in acetyl alpha tubulin level at 80 to 400 nM after 6 hrs by Western blot analysis	30901208
HEL	Function assay	100 mg/kg	5 days	Drug level in tumor of BALB/c nu mouse xenografted with HEL cells at 100 mg/kg/day, ip administered for 5 days and measured 1 hr post-last dose by LC-MS/MS analysis	30901208
Sf9	Function assay		30 secs	Inhibition of human recombinant N-terminal hexahistidine tagged JAK2 JH1 catalytic domain (835 to 1132 residues) expressed in baculovirus infected Sf9 cells using Tyr6 peptide as substrate incubated for 30 secs under shaking condition measured after 1 hr , IC50=0.0006μM	30981578
Sf21	Function assay		1 hr	Inhibition of recombinant human N-terminal epitope-tagged JAK2 (828 to 1132 residues) expressed in baculovirus infected Sf21 insect cells using EQEDEPEGDYFEWLE as substrate after 1 hr by homogeneous time-resolved fluorescence assay, IC50=0.0028μM	30981578
Sf9	Function assay		30 secs	Inhibition of human recombinant N-terminal hexahistidine tagged JAK1 JH1 catalytic domain (854 to 1154 residues) expressed in baculovirus infected Sf9 cells using Tyr6 peptide as substrate incubated for 30 secs under shaking condition measured after 1 hr , IC50=0.004μM	30981578
Sf9	Function assay			Binding affinity to human recombinant N-terminal hexahistidine tagged JAK2 JH1 catalytic domain (835 to 1132 residues) expressed in baculovirus infected Sf9 cells assessed as dissociation constant by surface plasmon resonance assay, Kd=0.0282μM	30981578
Sf9	Function assay		30 secs	Inhibition of human recombinant C-terminal hexahistidine tagged JAK3 JH1 catalytic domain (811 to 1124 residues) expressed in baculovirus infected Sf9 cells using Tyr6 peptide as substrate incubated for 30 secs under shaking condition measured after 1 hr , IC50=0.051μM	30981578
HEL	Antiproliferative assay		3 days	Antiproliferative activity against HEL cells harboring JAK2 V617F mutant measured after 3 days by CCK8 assay, IC50=7.639μM	30981578
insect cells	Function assay		10 mins	Inhibition of recombinant human N-terminal GST-tagged JAK1 (866 to 1154 residues) expressed in insect cells using FITC-labeled C6-KKHTDDGYMPMSPGVA-NH peptide as substrate after 10 mins in presence of 5 mM ATP by caliper mobility shift assay, IC50=0.02μM	32297743
insect cells	Function assay		10 mins	Inhibition of recombinant human N-terminal GST-tagged JAK2 (831 to 1132 residues) expressed in insect cells using 5FAM-labeled GEEPLYWSFPAKKK-NH2 peptide as substrate after 10 mins in presence of 5 mM ATP by caliper mobility shift assay, IC50=0.02μM	32297743
BAF3	Cytotoxicity assay		48 hrs	Cytotoxicity against mouse BAF3 cells expressing JAK2 V617F mutant after 48 hrs by CellTiterGlo assay, IC50=0.126μM	ChEMBL
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Chemische informatie, Opslag en Stabiliteit

Moleculair gewicht	404.36	Formule	C₁₇H₁₈N₆.H₃O₄P	Opslag (Vanaf de ontvangstdatum)	3 jaar-20°Cpoeder
CAS-nr.	1092939-17-7	--		Opslag van stamoplossingen	1 jaar-80°Cin oplosmiddel 1 maand-20°Cin oplosmiddel
Synoniemen	INCB018424, INC424			Smiles	C1CCC(C1)C(CC#N)N2C=C(C=N2)C3=C4C=CNC4=NC=N3.OP(=O)(O)O

Oplosbaarheid

In vitro
Batch:

DMSO : 81 mg/mL (200.31 mM)
(Met vocht verontreinigde DMSO kan de oplosbaarheid verminderen. Gebruik verse, watervrije DMSO.)

Ethanol : 9 mg/mL

Water : Insoluble

Molariteitscalculator

Massa	Concentratie	Volume	Moleculair gewicht
=	×	×

Verdunningscalculator Moleculair gewicht calculator

In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Gevalideerd door Selleck labs. Mocht u aanpassingen aan deze formulering nodig hebben, neem dan contact op met ons verkoopteam voor aangepaste tests.	6.000mg/ml (14.84mM)	Taking the 1 mL working solution as an example, add 50 μL of 120 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.

In vivo Formuleringscalculator (Heldere oplossing)

Stap 1: Voer de onderstaande informatie in (Aanbevolen: Een extra dier voor het geval van verlies tijdens het experiment)

Dosering: mg/kg Gemiddeld gewicht van dieren: g Doseervolume per dier:μL Aantal dieren:

Stap 2: Voer de in vivo formulering in (Dit is alleen de calculator, geen formulering. Neem eerst contact met ons op als er geen in vivo formulering is in het gedeelte Oplosbaarheid.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Berekeningsresultaten:

Werkconcentratie: mg/ml;

Methode voor het bereiden van DMSO-mastervloeistof: mg geneesmiddel vooraf opgelost in μL DMSO ( Concentratie mastervloeistof mg/mL, Neem eerst contact met ons op als de concentratie de DMSO-oplosbaarheid van de partij geneesmiddel overschrijdt. )

Methode voor het bereiden van in vivo formulering: Neem μL DMSO mastervloeistof, voeg vervolgens toeμL PEG300, mengen en helder maken, voeg vervolgens toeμL Tween 80, mengen en helder maken, voeg vervolgens toe μL ddH₂O, mengen en helder maken.

Methode voor het bereiden van in vivo formulering: Neem μL DMSO mastervloeistof, voeg vervolgens toe μL Maïsolie, mengen en helder maken.

Opmerking: 1. Zorg ervoor dat de vloeistof helder is voordat u het volgende oplosmiddel toevoegt.
2. Zorg ervoor dat u het/de oplosmiddel(en) in de juiste volgorde toevoegt. U moet ervoor zorgen dat de verkregen oplossing, bij de vorige toevoeging, een heldere oplossing is voordat u verdergaat met het toevoegen van het volgende oplosmiddel. Fysische methoden zoals vortexen, echografie of een warmwaterbad kunnen worden gebruikt om het oplossen te bevorderen.

Werkingsmechanisme

Targets/IC₅₀/Ki	JAK2 (Cell-free assay) 2.8 nM JAK1 (Cell-free assay) 3.3 nM
In vitro	INCB018424 remt krachtig en selectief JAK2V617F-gemedieerde signalering en proliferatie in Ba/F3-cellen en HEL-cellen. INCB018424 verhoogt de apoptose aanzienlijk op een dosisafhankelijke manier in Ba/F3-cellen. INCB018424 (64 nM) resulteert in een verdubbeling van cellen met gedepolariseerde mitochondria in Ba/F3-cellen. INCB018424 remt de proliferatie van erytroïde progenitorcellen van normale donoren en patiënten met polycythemia vera met een IC50 van respectievelijk 407 nM en 223 nM. INCB018424 toont een opmerkelijke potentie tegen erytroïde kolonieformatie met een IC50 van 67 nM.
Kinase Assay	Bindingstest
Kinase Assay	Recombinante eiwitten worden tot expressie gebracht met Sf21-cellen en baculovirusvectoren en gezuiverd met affiniteitschromatografie. JAK kinase assays gebruiken een homogene tijd-opgeloste fluorescentie assay met het peptidesubstraat (-EQEDEPEGDYFEWLE). Elke enzymreactie wordt uitgevoerd met Ruxolitinib of controle, JAK-enzym, 500 nM peptide, adenosine trifosfaat (ATP; 1mM), en 2% dimethylsulfoxide (DMSO) gedurende 1 uur. De 50% remmende concentratie (IC50) wordt berekend als de INCB018424-concentratie die nodig is voor de remming van 50% van het fluorescentiesignaal.
In vivo	INCB018424 (180 mg/kg, oraal, tweemaal daags) resulteert in een overlevingspercentage van meer dan 90% op dag 22 in een JAK2V617F-gestuurd muismodel. INCB018424 (180 mg/kg, oraal, tweemaal daags) vermindert splenomegalie en circulerende niveaus van inflammatoire cytokines aanzienlijk, en elimineert bij voorkeur neoplastische cellen, wat resulteert in een significant verlengde overleving zonder myelosuppressieve of immunosuppressieve effecten in een JAK2V617F-gestuurd muismodel. Het primaire eindpunt wordt bereikt bij 41,9% van de patiënten in de Ruxolitinib-groep in vergelijking met 0,7% in de placebogroep in de dubbelblinde studie van myelofibrose. Ruxolitinib resulteert in het behoud van de vermindering van het miltvolume en een verbetering van 50% of meer in de totale symptoomscore. In totaal heeft 28% van de patiënten in de Ruxolitinib (15 mg tweemaal daags) groep ten minste een 35% vermindering van het miltvolume in week 48 bij patiënten met myelofibrose, vergeleken met 0% in de groep die de best beschikbare therapie kreeg. De gemiddelde palpabele miltlengte is met 56% afgenomen met Ruxolitinib, maar met 4% toegenomen met de best beschikbare therapie in week 48. Patiënten in de ruxolitinib-groep vertonen een verbetering in de algemene kwaliteit van leven en een vermindering van symptomen geassocieerd met myelofibrose.
Referenties	[1] https://pubmed.ncbi.nlm.nih.gov/20130243/ [2] https://pubmed.ncbi.nlm.nih.gov/22375971/ [3] https://pubmed.ncbi.nlm.nih.gov/22375970/ [4] https://pubmed.ncbi.nlm.nih.gov/20130243/

Informatie klinische proef

(gegevens van https://clinicaltrials.gov, bijgewerkt op 2024-05-22)

NCT-nummer	Rekrutering	Aandoeningen	Sponsor/Medewerkers	Startdatum	Fasen
NCT06310304	Active not recruiting	Healthy Participants	Incyte Corporation	March 26 2024	Phase 1
NCT02596347	Completed	Chronic Beryllium Disease (CBD)\|Beryllium Sensitization (BeS)	National Jewish Health	April 2015	--

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